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PubMed1989

Increased dosage of diphtheria toxoid for basic immunization of adults.

Basic immunization of adults with increased dosages of a diphtheria toxoid vaccine (2100 flocculation units (Lf)/mg) was evaluated. Three injections of 7.5 Lf or 15 Lf diphtheria toxoid were given to 243 adults who had a history of no more than one previous vaccine injection. Systemic reactions were rare in both groups. Following the first two injections, local reactions (greater than 5 cm) were observed in 6-14% of the adults. After the third injection, 35% of adults in the 15 Lf group reported a local reaction (greater than 5 cm) compared to 11% in the 7.5 Lf group (p less than 0.001). The 15 Lf dose elicited a better antitoxin response than the 7.5 Lf dose. In a seronegative subgroup including 65 vaccinees who showed no booster response to the first vaccination, 79% had a postvaccination titer of greater than or equal to 0.1 IU/ml and 28% a titer of greater than or equal to IU/ml after the third injection of 7.5 Lf. The corresponding numbers in the 15 Lf group were 94% and 44%, respectively. The study demonstrates that 7.5 Lf and 15 Lf diphtheria toxoid of high purity can safely be given to adults for basic immunization. The higher dose is more immunogenic but local reactions increase after the third injection.

Adult↗
PubMed1999

Does concomitant injection of a combined diphtheria-tetanus-acellular pertussis - hepatitis B virus - inactivated polio virus vaccine influence the reactogenicity and immunogenicity of commercial Haemophilus influenzae type b conjugate vaccines?

UNLABELLED: Does concomitant injection of a combined diphtheria-tetanus-acellular pertussis - hepatitis B virus - inactivated polio virus vaccine influence the reactogenicity and immunogenicity of commercial Haemophilus influenzae type b conjugate vaccines? The immunogenic responses and local reactions to four Haemophilus influenzae type b (Hib) conjugate vaccines licensed for primary immunisation (Hiberix, ActHib, Pedvax, HibTITER) when administered concomitantly but in the opposite thigh with a candidate diphtheria-tetanus-acellular pertussis-hepatitis B-inactivated poliovirus vaccine were studied in 549 healthy infants at 3, 4.5 and 6 months of age. Local reactions were mild, but different between the four groups, a tetanus conjugate Hib vaccine showing the fewest reactions. All local reactions resolved without sequelae. There was no apparent general reaction. The immunogenic response was similar with all four vaccines, geometric mean concentrations ranging from 4.95 to 7.2 microg/ml. All but one subject had antipolyribosylribitol phosphate polysaccharide antibody titres > or =0.15 microg/ml, and 88.0% to 96% achieved high titres (>1.0 microg/ml) generally associated with long-term protection against Hib disease. CONCLUSION: There does not appear to be any interference with the immune response when current commercial Haemophilus influenzae type b conjugate vaccines are concomitantly administered with a candidate diphtheria-tetanus-acellular pertussis-hepatitis B-inactivated polio virus vaccine as separate injections.

Antibodies, Bacterial↗
PubMed2004

Alternative diphtheria, tetanus and whooping cough immunization schedule to evoke a Th2 tetanus and a Th1 pertussis immune response.

In a previous study, using BALB/c mice, we found that while diphtheria (D), tetanus (T) and whooping cough (Pw, whole-cell Bordetella pertussis) immunization induces a Th1/Th2 tetanus response and memory T cells able to proliferate in response to in vitro stimulation with B. pertussis, DTPa immunization induces a Th2 tetanus immune response and no memory T cells that recognize B. pertussis as stimulus. Considering that a pro-inflammatory cytokine production is not necessary for protection against tetanus and therefore should be avoided, an alternative DTP immunization schedule with minimal Pw exposure was assessed in order to obtain a Th2 tetanus response and a Th1 pertussis response. BALB/c mice were primed with DT vaccine at day 0, with Pw vaccine at day 14 and boosted with DTPa vaccine at days 21 and 28. A control group was inoculated with saline. Antibodies against B. pertussis surface antigens, tetanus and diphtheria toxoids were produced by mice. Spleen cells stimulated in vitro with B. pertussis produced IL-6 and IFNgamma. Only IL-5 was produced by cells in response to tetanus toxoid stimulation. These results are in line with the low IgG1/IgG2a ratio for pertussis antibodies compared with those corresponding to tetanus and diphtheria. The immunization protocol presented herein succeeded in producing tetanus and pertussis immune responses of Th2 and Th1 type, respectively. In contrast to previous results obtained with DTPw immunization, no IL-12 production was observed. Our findings provide direct evidence that an immunization protocol with an interval of 14 days between DT and Pw primings, followed by DTPa boosters, can induce appropriate immune responses against DTP vaccine antigens.

Animals↗
PubMed1992

Safety and immunogenicity of acellular diphtheria-tetanus-pertussis and Haemophilus conjugate vaccines given in combination or at separate injection sites.

This prospective, double-blind, randomized trial compared the immunogenicity and reactogenicity of acellular diphtheria-tetanus-pertussis vaccine and Haemophilus influenzae type b conjugate vaccine-diphtheria toxoid conjugate, given at separate injection sites or at a single site, in 79 children 18 months of age who had received three prior immunizing doses of whole-cell diphtheria-tetanus-pertussis vaccine. No significant differences were observed.

Antibodies, Bacterial↗
PubMed1999

Study on the use of an enzyme-linked immunosorbent assay in determining human antibodies to diphtheria toxin as compared with a reference toxin neutralization assay.

Serum samples from 156 Greek persons were assessed by an IgG-specific enzyme-linked immunosorbent assay (ELISA) and a reference tissue culture toxin-neutralization (TN) assay for the quantitation of diphtheria toxin antibodies. By the reference method, 7.7% of the persons were susceptible to diphtheria (antitoxin < 0.01 IU/ml), 28.8% had basic protection (antitoxin 0.01-0.09 IU/ml) and 63.5% were fully protective (antitoxin > or = 0.1 IU/ ml), while the corresponding figures were 17.9, 36.5 and 45.5% when they were tested by the immunoassay. None of the samples been susceptible by the TN assay were found to have some protection when tested by ELISA. However, three (6.7%) of the 45 samples showing a basic protection with TN, were fully protective when titrated by the immunoassay. In addition, 31 (31.3%) of the 99 samples been fully protective by the bioassay, were found to be either basically protective or susceptible by means of the ELISA. Overall, validity features of the immunoassay were: sensitivity 68.7%, specificity 94.7%, positive predictive value 95.8% and negative predictive value 63.5%. The ELISA tested in our study could be used to determine diphtheria antitoxin in individuals needed a booster immunization (susceptible or basic protective samples), although it might falsely include in the above categories samples that are within the fully protective levels of antibodies.

Adolescent↗
PubMed1998

Non-toxigenic Corynebacterium diphtheriae: two cases and review of the literature.

Non-toxigenic Corynebacterium diphtheriae infections are being reported with increasing frequency. We present two cases of C. diphtheriae endocarditis requiring early valve replacement. Both cases were complicated by cerebral embolic phenomena and pseudoaneurysm formation in lower limb arterial vessels. Non-toxigenic C. diphtheriae septicaemia must be excluded when 'diphtheroids' are isolated from blood cultures.

Adolescent↗
PubMed2003

Effect of infant immunisation with meningococcus serogroup C-CRM(197) conjugate vaccine on diphtheria immunity and reactogenicity in pre-school aged children.

The majority of Men C conjugate vaccines given in the UK use CRM(197), a mutant diphtheria toxoid, as their protein carrier. We studied the effects of prior immunisation with Men C-CRM(197) conjugate vaccine on immunity to diphtheria in 193 children before and after a booster dose of Men C at 4 years. Baseline diphtheria antibodies were higher in children given four previous doses of Men C (P<0.0001) and tended to be higher following boosting in those who had received three or four doses. This enhanced immunity was not associated with increased reactogenicity.

Child, Preschool↗
PubMed1999

Immunogenicity of IRIV- versus alum-adjuvanted diphtheria and tetanus toxoid vaccines in influenza primed mice.

The immunogenicity and protectivity of two different toxoid vaccines were compared in mice. In one formulation, toxoids (diphtheria or tetanus) were adsorbed to alumoxid, whereas in the other formulation the toxoids were crosslinked to immunopotentiating reconstituted influenza virosomes (IRIVs). A preimmunization with influenza antigens is necessary for a good anti-toxoid antibody response when the IRIV formulation was administered. After two immunizations with the IRIV- or alum-based vaccines, the IRIV-based formulation induced a higher humoral immune response than toxoids adsorbed to alum. Using an in vitro test, diphtheria toxin neutralizing antibodies were tested. Di-IRIV induced a significantly (p = 0.002) higher titer of diphtheria toxin neutralizing antibodies than Di-alum. Tetanus challenge experiments showed, that the IRIV-based tetanus vaccine induced a threefold higher titer of protective antibodies than the tetanus toxoid adsorbed to alum. Therefore, the IRIV-based formulations appeared to be superior to the alum-based vaccines in terms of immunogenicity and protectivity.

Adjuvants, Immunologic↗
PubMed1999

Evaluation of the effectiveness of vaccination against diphtheria among medical staff of the chair and the Jagiellonian University Hospital of Infectious Diseases in Cracow, Poland. Some epidemiological observations.

In Poland, twenty-five cases of diphtheria, mostly among adults (including five foreigners) in the region adjacent to the eastern border were registered in the years 1992-1996. In 1993-1994, because of the threat of diphtheria infection, 41 healthy members of the medical staff of the University Hospital of Infectious Diseases in Kraków, mainly women aged 25-57 years were immunized with Td and d vaccines with a 6-month interval between courses. Prior to vaccination, 17(41%) individuals showed a lack of diphtheria antibodies as determined with a passive haemagglutination test. 14 of them were aged 30-49 years. Six months after the 2nd dose of vaccine, all seronegative persons had developed a protective level of antibodies. Moreover, all vaccinated subjects showed a statistically significant increase of the antibody titre between the first and last serological examinations.

Adult↗
PubMed1972

Diphtheria infection in North West Canada, 1969, 1970 and 1971.

In three years, Corynebacterium diphtheriae was isolated from 1238 people, consisting of 820 North American Indians or Metis, 318 people of Caucasian origin, 97 Eskimos and 3 Asiatic Indians. Diphtheria infection of the throat, nose, ears and skin was common in the North American Indian and Metis people, but rarely caused severe symptoms. The infection occurred less often in white people but was more serious; of 27 cases of toxic respiratory diphtheria, 25 were white people. The public health significance of the endemic infection of the North American Indian and Metis people is discussed.

Adolescent↗
PubMed1984

Population studies of diphtheria immunity using antitoxin radioimmunoassay.

A quantitative method for testing serum diphtheria antitoxin levels was set up using a diphtheria antitoxin radioimmunoassay (RIA). The results of this RIA correlated well with the Schick test in 554 subjects and with intradermal neutralization tests in guinea-pigs in a small group of subjects. The RIA was suitable for use on blood collected by fingerprick on to a disc of standard chromatography paper. These discs could be stored at room temperature for at least 1 month. If storage for more than 6 months was required -20 degrees C was found to be better. Experience with this RIA in a total of 2349 subjects indicated that it is more accurate, rapid and less costly than Schick testing. The RIA should prove to be the preferred method for testing diphtheria immunity in population surveys.

Adolescent↗
PubMed1989

Immunity against diphtheria in adults in Poland.

The diphtheria immunity status was determined with the passive haemagglutination technique in 503 sera of 10-90-year-old persons from Warsaw and Olsztyn Provinces. Donors of sera were students, teachers, pregnant women, employees of industry and medical service. The immunity was highest (90% of titers 0.1 IU/ml or higher) in persons below 20 years of age and in persons above 60 years of age (55%). Between these two groups, gaps in immunity exist, the proportion of those immune varying from 36-50% in the 20- 60-year-old groups. Since a large pool of susceptible persons creates an epidemic potential it was suggested that the adult type of tetanus-diphtheria toxoid (Td) should be introduced into the routine immunization schedule for high risk groups. These groups might include professional or age groups who are vulnerable to reintroduction of virulent Corynebacterium diphtheriae such as kindergarten and creches personnel, teachers, students, military service personnel and persons travelling to developing countries.

Adolescent↗
PubMed1994

Measurement of diphtheria and tetanus antitoxin in blood samples collected on filter paper disks.

Large-scale surveys of immunity to vaccine-preventable diseases may be limited by the inconvenience and expense involved in collection of blood by venepuncture. An alternative method of collecting blood on filter paper for measurement of immunity to diphtheria and tetanus is described. The precut filter disks (Elisadiscs), originally developed for serological diagnosis of disease in pigs, have advantages over previously described methods in that they allow safe handling of minimal volumes of blood (5 microliters) which can be conveniently quantified. To compare values obtained by venepuncture and fingerprick, paired samples were collected from 60 subjects and diphtheria and tetanus antitoxin concentrations were measured by ELISA. There was no significant difference detected between samples collected by the two methods. The results suggest that Elisadiscs are a reliable alternative to venepuncture for monitoring immunity to diphtheria and tetanus and would be useful for sample collection in remote areas and from children.

Adult↗
PubMed2002

The amino acid requirements of Corynebacterium diphtheriae PW 8 substrain CN 2000.

AIMS: To determine the amino acid requirement and utilization pattern of Corynebacterium diphtheriae during growth and toxin production. METHODS AND RESULTS: Comparing across different batches of beef-based media, the growth and toxin yield were correlated significantly with nine of the amino acids. The amino acid utilization pattern during growth of C. diphtheriae further showed that only four of the nine amino acids, namely cystine, histidine, aspartate and methionine, were critical for growth of the vaccine strain. Further investigations using synthetic media with combinations of amino acid supplements demonstrated that among the four, cystine was the most growth limiting. CONCLUSIONS: Only certain amino acids are critical for growth and toxin production by C. diphtheriae, cystine being the single most important. SIGNIFICANCE AND IMPACT OF THE STUDY: Owing to the potential threat from Bovine Spongiform Encephalopathy (BSE), a need is recognized by vaccine manufacturers to substitute beef-based production media. An understanding of the specific amino acid requirements would help to develop and optimize alternative production media.

Amino Acids↗
PubMed1994

[Toxic diphtheria caused by Corynebacterium ulcerans].

A 42-year-old woman with bronchial asthma since childhood was admitted to hospital because of severe dyspnoea. Emergency bronchoscopic intubation had to be performed for life-threatening inspiratory and expiratory stridor. This demonstrated that the larynx was covered by a dirty-grey membrane and the vocal-cord gap was narrowed to a mere slit. As laryngeal diphtheria was suspected 2000 IU/kg diphtheria antitoxin was administered together with 1 mega U penicillin G four times daily intravenously. On the same day, Corynebacterium ulcerans, a very rare cause of diphtheria, was isolated from a coughed-up piece of the membrane. Toxic, massive swelling of the lymph nodes and soft tissues of the neck necessitated maintenance of an open upper airway by intubation and (later) tracheostomy for 41 days. A week later the patient was discharged without any permanent defect.

Adult↗
PubMed1981

[Level of immunity against diphtheria among the personnel of a Hamburg hospital and in the normal population (author's transl)].

Diphtheria antitoxin titres were measured by passive haemagglutination in a total of 4542 blood samples obtained between the end of 1980 and beginning of 1981 from staff at Hamburg hospitals and from randomly selected age-matched persons (1219). Among the hospital personnel 67.5% had no protective diphtheria antitoxin (less than or equal to 0.11 IU/ml), compared with 52% among the control group. Among those aged 15-24 years, 60.2% of staff were unprotected, compared with 46.6% of the control population. The geometric mean value of the diphtheria antitoxins also gave a worse immunity level for hospital personnel (0.08 IU/ml) than the control population (0.14 IU/ml). While geometric mean titre values of hospital staff beyond the age of 45 fell nearly continuously, those in the control population rose with increasing age.

Adolescent↗
PubMed1983

Nucleotide sequence of the structural gene for diphtheria toxin carried by corynebacteriophage beta.

A 1,942-base-pair DNA segment encoding the structural gene for diphtheria toxin was sequenced, and the primary structure of the toxin was deduced. Restriction enzyme fragments corresponding to nontoxic or hypotoxic peptides of the toxin were isolated from corynebacteriophage beta and cloned into Escherichia coli on plasmid pBR322, and the sequence was determined. The mature toxin molecule deduced from the sequence has 535 amino acid residues and a molecular weight of 58,342. The deduced sequence for the fragment A moiety was the same as that determined at the protein level, except for a single serine residue, which had been mispositioned in the earlier study. Several differences were noted with respect to the partial sequence data available on the fragment B moiety, some or all of which may reflect genetic variations among populations of corynephages carrying the toxin gene. The DNA sequence predicts a 25-residue leader peptide preceding the mature protein, which is presumably involved in secretion of the toxin from lysogenized Corynebacterium diphtheriae. We infer that initiation of translation probably occurs at a GTG codon (codon -25). Cloned restriction fragments containing sequences for the amino-terminal region of toxin, together with 5' flanking regions, were expressed in E. coli. Toxin-related peptides were synthesized and secreted into the periplasmic space. These results provide a basis for applying recombinant DNA methods to the study of diphtheria toxin and for producing novel, genetically altered forms of the toxin suited to the construction of new classes of immunotoxins.

Amino Acid Sequence↗
PubMed1988

Interleukin 2-diphtheria toxin fusion protein can abolish cell-mediated immunity in vivo.

De novo expression of the interleukin 2 receptor (IL-2R) is a critical and pivotal event in initiation of an immune response. Targeting the low-affinity IL-2-binding p55 subunit of the high-affinity IL-2R with the rat anti-mouse IgM monoclonal antibody M7/20 suppresses a variety of T-cell-mediated reactions, including transplant rejection, autoimmunity, and delayed-type hypersensitivity (DTH). A hybrid IL-2-toxin gene was constructed from the diphtheria toxin gene by replacing the DNA encoding the diphtheria toxin receptor-binding domain with the DNA encoding the receptor-binding domain of IL-2, and the fusion protein encoded by the hybrid gene was expressed in Escherichia coli [Williams, D.P., Parker, K., Bacha, P., Bishai, W., Borowski, M., Genbauffe, F., Strom, T.B. & Murphy, J.R. (1987) Protein Eng. 1, 493-498]. We examined the action of the chimeric IL-2-toxin fusion protein on an in vivo T-cell mediated response, DTH. The IL-2-toxin fusion protein was found to be a potent immunosuppressive agent. Treatment of mice with the IL-2-toxin blocks DTH and prevents expansion of IL-2R+ T cells. Indeed, IL-2-toxin treatment targets IL-2R+ T cells in vivo and is shown to selectively eliminate their appearance in draining lymph nodes. DTH suppression was observed even in mice possessing high titers of antibodies to diphtheria toxoid.

Animals↗